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J. Biol. Chem., Vol. 277, Issue 43, 41213-41219, October 25, 2002
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From the To understand the role of eicosanoids in
angiogenesis, we have studied the effect of lipoxygenase metabolites of
arachidonic acid on human microvascular endothelial cell (HMVEC) DNA
synthesis. Among the various lipoxygenase metabolites of arachidonic
acid tested, 5(S)-hydroxyeicosatetraenoic acid
(5(S)-HETE) induced DNA synthesis in HMVEC.
5(S)-HETE also stimulated Jak-2, STAT-1, and STAT-3
tyrosine phosphorylation and STAT-3-DNA binding activity. Tyrphostin
AG490, a specific inhibitor of Jak-2, significantly reduced tyrosine
phosphorylation and DNA binding activity of STAT-3 and DNA synthesis
induced by 5(S)-HETE. In addition, 5(S)-HETE stimulated phosphatidylinositol 3-kinase (PI3-kinase) activity and
phosphorylation of its downstream targets Akt, p70S6K, and 4E-BP1 and
their effector molecules ribosomal protein S6 and eIF4E. LY294002 and
rapamycin, potent inhibitors of PI3-kinase and mTOR, respectively, also blocked the DNA synthesis induced by
5(S)-HETE. Interestingly, AG490 attenuated
5(S)-HETE-induced PI3-kinase activity and phosphorylation
of Akt, p70S6K, ribosomal protein S6, 4E-BP1, and eIF4E.
5(S)-HETE induced the expression of basic fibroblast growth
factor 2 (bFGF-2) in a Jak-2- and
PI3-kinase-dependent manner. In addition, a neutralizing
anti-bFGF-2 antibody completely blocked 5(S)-HETE-induced
DNA synthesis in HMVEC. Together these results suggest that
5(S)-HETE stimulates HMVEC growth via Jak-2- and
PI3-kinase-dependent induction of expression of bFGF-2.
These findings also reveal a cross-talk between Jak-2 and PI3-kinase in
response to 5(S)-HETE in HMVEC.
5(S)-Hydroxyeicosatetraenoic Acid Stimulates DNA
Synthesis in Human Microvascular Endothelial Cells via Activation of
Jak/STAT and Phosphatidylinositol 3-Kinase/Akt Signaling, Leading to
Induction of Expression of Basic Fibroblast Growth Factor 2*
§,§,, and¶
Department of Physiology and ¶ Center
for Vascular Biology, University of Tennessee Health Science Center,
Memphis, Tennessee 38163
*
This work was supported by National Institutes of Health
Grant HL64165 (to G. N. R.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of
Physiology, University of Tennessee Health Science Center, 894 Union Ave., Memphis, TN 38163. Tel.: 901-448-7321; Fax: 901-448-7126; E-mail:
grao@physio1.utmem.edu.
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